Table Of ContentALKALINE HYDROLYSIS OF EXPLOSIVES
by
Catherine Elizabeth VanEngelen
A dissertation submitted in partial fulfillment
of the requirements for the degree
of
Doctor of Philosophy
in
Engineering
MONTANA STATE UNIVERSITY
Bozeman, Montana
January 2010
©COPYRIGHT
by
Catherine Elizabeth VanEngelen
2010
All Rights Reserved
ii
APPROVAL
of a dissertation submitted by
Catherine Elizabeth VanEngelen
This dissertation has been read by each member of the dissertation committee and
has been found to be satisfactory regarding content, English usage, format, citation,
bibliographic style, and consistency, and is ready for submission to the Division of
Graduate Education.
Dr. Brent M. Peyton
Approved for the Department of Chemical and Biological Engineering
Dr. Ron W. Larsen
Approved for the Division of Graduate Education
Dr. Carl A. Fox
iii
STATEMENT OF PERMISSION TO USE
In presenting this dissertation in partial fulfillment of the requirements for a
doctoral degree at Montana State University, I agree that the Library shall make it
available to borrowers under rules of the Library. I further agree that copying of this
dissertation is allowable only for scholarly purposes, consistent with “fair use” as
prescribed in the U.S. Copyright Law. Requests for extensive copying or reproduction of
this dissertation should be referred to ProQuest Information and Learning, 300 North
Zeeb Road, Ann Arbor, Michigan 48106, to whom I have granted “the exclusive right to
reproduce and distribute my dissertation in and from microform along with the non-
exclusive right to reproduce and distribute my abstract in any format in whole or in part.”
Catherine Elizabeth VanEngelen
January 2010
iv
DEDICATION
This work is dedicated to my parents, Patricia and Daniel Albaugh, and to my
husband, Michael VanEngelen. Only their loving support and encouragement has made
this possible.
v
ACKNOWLEDGEMENTS
Financial support for this research was provided by: the Thermal Biology Institute
at MSU (TBI grant #NNG04GR46G), the Inland Northwest Research Alliance (INRA
Student Fellowship), the Center for Biofilm Engineering at MSU, the College of
Engineering at MSU (Benjamin Fellowship), and the Radford Army Ammunition Plant,
Radford, VA.
I would also like to gratefully acknowledge the following people for their help
and guidance throughout my time at Montana State: Shelley Thomas, Jill Newman, John
Neuman, Ann Willis, John and Flower Aston, Melinda Clark, Angela and Dennis
O’Brien, Chris and Angie Albaugh, Mary and Dean Lutz, Patrick Albaugh, John and
Susan VanEngelen, Tamara and Rich Cullen, Jen and Tim Traynor, Kristen and Chip
George, members of the Peyton Research Group, Ron Larsen, and my committee: Ross
Carlson, Robin Gerlach, Anne Camper, and Brent Peyton.
vi
TABLE OF CONTENTS
1. INTRODUCTION .................................................................................................... 1
2,4,6-Trinitrotoluene (TNT)...................................................................................... 1
Abiotic Treatments............................................................................................... 2
Biological Treatments .......................................................................................... 5
Nitrocellulose (NC) ................................................................................................. 11
Abiotic Treatments ............................................................................................ 13
Biological Treatments ........................................................................................ 15
Combined Hydrolysis and Biological Treatments ....................................... 16
Summary ................................................................................................................. 18
References ............................................................................................................... 19
2. ENRICHMENT FOR A TNT-DEGRADING MICROORGANISM .................... 24
Background ............................................................................................................. 24
Materials and Methods ............................................................................................ 24
Results and Discussion ........................................................................................... 26
Conclusions ............................................................................................................. 32
References ............................................................................................................... 33
3. ALKALINE HYDROLYSIS OF TNT ................................................................... 39
Introduction ............................................................................................................. 39
Materials and Methods ............................................................................................ 41
Chemicals ........................................................................................................... 41
Hydrolysis Experiments ..................................................................................... 41
Sampling and Storage ........................................................................................ 42
Analyses ............................................................................................................. 42
Radiolabeled TNT Experiments ........................................................................ 43
Biological Screening .......................................................................................... 43
Results and Discussion ........................................................................................... 44
Kinetics of Hydrolysis ....................................................................................... 47
Hydrolysis Products ........................................................................................... 53
Radiolabeled 14C-TNT Experiments .................................................................. 55
Biological Degradation of TNT Hydrolysate .................................................... 58
Conclusions ............................................................................................................. 60
References ............................................................................................................... 62
vii
TABLE OF CONTENTS – CONTINUED
4. NC TREATABILITY STUDY ............................................................................... 68
Background ............................................................................................................. 68
Materials and Methods ............................................................................................ 69
Results and Discussion ........................................................................................... 71
Conclusions ............................................................................................................. 79
Recommendations ................................................................................................... 81
References ............................................................................................................... 83
5. ALKALINE HYDROLYSIS OF NC ..................................................................... 85
Introduction ............................................................................................................. 85
Materials and Methods ............................................................................................ 87
Results and Discussion ........................................................................................... 89
Hydrolysis of NC ............................................................................................... 89
Biological Degradation of NC Hydrolysate ....................................................... 95
Conclusions ............................................................................................................. 98
References ............................................................................................................... 99
6. SUMMARY AND FUTURE WORK .................................................................. 101
TNT ....................................................................................................................... 101
NC ......................................................................................................................... 103
APPENDICES ............................................................................................................ 104
APPENDIX A: Consensus Sequence for Anoxybacillus kualowohkensis ............ 105
APPENDIX B: Raw Data for TNT/TNB Experiments ........................................ 107
APPENDIX C: Raw Data for 14C-TNT Experiments ........................................... 112
APPENDIX D: Raw Data for Biological Degradation of TNT Hydrolysate ....... 119
APPENDIX E: Raw Data for TNT Hydrolysis Experiments ............................... 121
APPENDIX F: HPLC 1090 Chromatogram: “10 Mix” Standards ....................... 134
APPENDIX G: HPLC 1090 Chromatogram: “14 Mix” Standards ...................... 136
APPENDIX H: Raw Data for NC Treatability Study ........................................... 138
APPENDIX I: Raw Data for NC Hydrolysis Experiments .................................. 142
APPENDIX J: Raw Data for NC Hydrolysate Biological Screening .................. 147
viii
LIST OF TABLES
Table ............................................................................................................................... Page
1. YNP spring samples ........................................................................................................ 25
2. Comparison of molar TNT and hydroxide concentrations at various
pH values ....................................................................................................................... 45
3. Comparison of pseudo-first-order kinetic rate constants, k’ (h-1) ................................... 50
4. Composition for NC experiments at both room temperature (RT)
and 60°C .......................................................................................................................... 69
5. Denitrification rates (mg NO / h · mg biomass) for
x
WWTP culture spiked with NC hydrolysate .................................................................. 97
ix
LIST OF FIGURES
Figure .............................................................................................................................. Page
1. Chemical structure of TNT ............................................................................................... 1
2. Pathways of biological TNT transformation..................................................................... 7
3. Chemical structure of NC ............................................................................................... 12
4. Biological treatment and controls for 16mg/L TNT degradation
with 1g/L yeast extract at 60°C and pH 9 ....................................................................... 27
5. Protein production for 16mg/L TNT degradation and 1g/L yeast
extract at 60°C and pH 9 ................................................................................................. 28
6. Occurrence of TNB as the abiotic product of 16mg/L TNT
degradation at 60°C and pH 9 ......................................................................................... 29
7. Chemical structure of TNB ............................................................................................. 29
8. Effect of pH and temperature on TNT concentration ..................................................... 46
9. TNT degradation with production of nitrite and nitrate at pH 9 ..................................... 47
10. TNT degradation with production of nitrite and nitrate at pH 10 ................................. 48
11. TNT degradation with production of nitrite and nitrate at pH 11 ................................. 48
12. Linearized Arrhenius plot for determination of activation energy
of the alkaline hydrolysis of TNT ................................................................................. 52
13. Initial distribution of 14C -TNT and corresponding HPLC
chromatogram ............................................................................................................... 56
14. Day 1 distribution of 14C from TNT treated at pH 10 and 60°C
and corresponding HPLC chromatogram ..................................................................... 57
15. Day 9 distribution of 14C from TNT treated at pH 10 and 60°C
and corresponding HPLC chromatogram ..................................................................... 58
16. Growth of WWTP denitrifying culture on TNT hydrolysate ....................................... 59
Description:ALKALINE HYDROLYSIS OF EXPLOSIVES by. Catherine Elizabeth VanEngelen. A dissertation submitted in partial fulfillment of the requirements for the degree of. Doctor of Philosophy in. Engineering. MONTANA STATE UNIVERSITY. Bozeman, Montana. January 2010
